The objective of the present study was to evaluate the effects of ghrelin on the concentrations of estrogen (E(2)) and progesterone (P(4)) in serum and the mRNA expression of estrogen receptor beta (ER(β)) and progesterone receptor (PR(A+B)) in ovary in rats during estrous cycle. Adult female Sprague Dawley rats were intracerebroventricularly (i.c.v.) injected with 3 nmol ghrelin during the estrous cycle, and sacrificed 15 min later. Blood samples and ovaries were collected. The concentrations of serum E(2) and P(4) were measured by radioimmunoassay, while the amount of ER(β) and PR(A+B) mRNA was assessed by real-time quantitative PCR. Our studies showed that ghrelin could significantly reduce the serum concentration of E(2) throughout the estrous cycle (P < 0.05), the serum level of P(4) (P < 0.05), and the amount of ER(β) mRNA during metestrus (P < 0.05). Meanwhile, the amount of PR(A+B) mRNA was only reduced during diestrus (P < 0.05). Overall, our present findings provide the first evidence that i.c.v. injection of ghrelin could reduce the serum concentration of E(2) and P(4) and the level of ER(β) and PR(A+B) mRNA expression, supporting the role of ghrelin in reproduction.